Pharmacokinetics and blood-brain barrier penetration of a novel salidroside derivative pOBz in rats北大核心CSCD

Aim To develop a ultra-high performance liquid chromatography electrospray-ionization tandem mass spectrometry ( UPLC-MS/MS ) method for the simultaneous determination of salidroside derivative pOBz in rat plasma and brain tissue, and to study the pharmacokinetic profile and penetration of the blood-brain barrier in rats after a single dose intravenous administration of pOBz. Methods SD rats were administered pOBz at a dose of 50 mg • kg -1via tail vein, and plasma and brain tissue samples were collected at different time points. UPLC-MS/MS method was used to determine the concentration of pOBz in plasma and brain tissue. The plasma concentration-time curve was fitted by DAS 2. 1. 1 software to calculate the pharmacokinetic parameters of pOBz. Results The linear regression equation of pOBz in plasma was: Y = 0.062 3X + 0.024 8 , r2=0.999, and the linear range was 0. 2 ~ 200 mg • L-1; the linearity regression equation in brain homogenate was : Y = 0.065 1X + 0.0317 , r2= 0.997, and the linear range was 0.2 ~ 200 mg • L -1. The half-life T1/2 of pOBz in plasma was 1.03 h,the area under the drug-time curve AUC0_twas 198.4 h • mg • L -1the mean residence time MRT0_twas 0.87 h, and the clearance rate CL was 0. 246 L • h-1• kg-1. 5 min after administration, pOBz could be detected in brain tissue at a concentration of 1.21 mg • L-1; it was not completely eliminated 3 h after administration. Conclusions The UPLC-MS/MS method is accurate and applicable for the pharmacokinetic study of pOBz in plasma and brain homogenate. pOBz can penetrate the blood-brain barrier and exert pharmacological effects in central nervous system. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

大黄对MCAO模型大鼠的神经保护作用

目的研究大黄提取物对MCAO模型大鼠的神经保护作用,并探讨其作用机制。方法SPF级雄性SD大鼠45只,随机分为sham组、MCAO组、MCAO+大黄组,线栓法制备MCAO模型,大黄提取物给药浓度为200 mg·kg^-1·d^-1,连续给药6 d,尼氏染色法检测尼氏体的表达,免疫荧光法检测大鼠缺血侧NeuN、NF200的表达,RT-qPCR法检测Egr1、Egr2、Egr4 mRNA表达,Western blot检测NGF及BDNF蛋白的表达。结果与MCAO组相比,大黄提取物作用后,尼氏染色法检测结果表明大黄提取物可以减少MCAO模型大鼠脑梗死面积,免疫荧光检测结果表明大黄提取物可以升高MCAO模型大鼠缺血侧NeuN、NF200的表达;经RT-qPCR检测,大黄提取物能够提高Egr1、Egr2、Egr4 mRNA基因表达;经Western blot检测,大黄提取物能够提高NGF及BDNF蛋白的表达。结论大黄提取物能够改善MCAO模型大鼠缺血侧神经元存活环境,具有神经保护作用。     

红景天苷调控PI3K/Akt信号通路对永久性脑缺血大鼠的保护作用

目的研究红景天苷(salidroside,Sal)通过调控PI3K/AKT信号通路对永久性大脑中动脉闭塞模型(pMCAO)大鼠的神经保护作用及其机制。方法健康成年雄性SPF级雄性SD大鼠70只,其中30只大鼠随机分为假手术组(sham组)、模型组(pMCAO组)、红景天苷给药组(pMCAO+Sal组),线栓法造模成功后,对sham组和pMCAO组大鼠腹腔注射生理盐水,pMCAO+Sal组按照50 mg·kg^-1·d^-1腹腔注射红景天苷(50 mg·kg^-1·d^-1),给药1 d。40只大鼠随机分为假手术组(sham组)、模型组(pMCAO组)、红景天苷给药组(pMCAO+Sal组)、抑制剂组(pMCAO+Sal+LY组);侧脑室注射PI3K抑制剂30 min后,制备pMCAO模型,1 h后腹腔注射红景天苷,给药1 d。免疫组织化学染色测定脑组织中NeuN表达,Western blot分析脑组织中NeuN蛋白、NF-κBp50核蛋白的表达,RT-qPCR检测IL-6和TNF-α的表达。结果与pMCAO组比较,红景天苷作用后,NeuN免疫荧光染色表达量明显增加,且能够促进NeuN蛋白,抑制NF-κB p50核蛋白及IL-6和TNF-αmRNA表达。经PI3K抑制剂LY294002作用后,红景天苷对上述指标作用不明显。结论红景天苷能够通过调控PI3K/AKT信号通路,抑制NF-κB p50核转录水平,进而抑制炎症因子,对pMCAO模型大鼠具有神经保护的作用。     

红景天苷对缺血性脑损伤大鼠的作用

目的 基于超高效液相色谱-电喷雾-串联质谱(UPLC-ESI-MS/MS)技术研究红景天苷(salidroside, Sal)的血脑屏障(blood brain barrier, BBB)穿透性,并通过网络药理学、分子对接技术和动物实验探讨Sal治疗缺血性脑卒中(ischemic stroke, IS)的作用靶点及机制。方法 利用UPLC-ESI-MS/MS技术研究Sal的BBB穿透性;运用多个数据库预测Sal的作用靶点和IS的疾病靶点,构建PPI网络并筛选核心靶点,进行GO和KEGG富集分析,并通过分子对接技术和动物实验进行验证。结果 Sal给药正常大鼠和MCAO大鼠后,血浆和脑组织中均检测到Sal原形和代谢物酪醇;网络药理学共筛选出Sal治疗IS的作用靶点191个,GO功能富集分析主要涉及蛋白水解、细胞迁移的正调控等生物过程;KEGG通路分析提示PI3K-Akt、MAPK、FOXO等信号通路在Sal治疗IS过程中发挥关键作用;分子对接结果显示,Sal与对接的核心靶点均有良好的结合能力;动物实验结果表明,Sal可以明显改善MCAO大鼠的神经功能损伤,明显增加缺血侧Nissl阳性细胞的...